Use of n-acetyl-d-glucosamine in the manufacture of pharmaceutical useful for treating cervical erosion

ABSTRACT

The present invention has disclosed the use of N-acetyl-D-glucosamine in the manufacture of a medicament for treating cervical erosion. N-acetyl-D-glucosamine can resist the field planting of microorganism and control microorganism infection, and improve the symptoms of local exudation, tissue inflammatory oedema and pain and the like.

TECHNICAL FIELD

[0001] The present invention relates to the use ofN-acetyl-D-glucosamine and pharmaceutical acceptable salts thereof inthe manufacture of a medicament for treating cervical erosion.

BACKGROUND ART

[0002] Cervical erosion is a common disease in gynecology, it is causedby superficial defect formed by squamous epithelium necrosis andabscission at cervix and vagina, or eversion of the mucosa at cervicalcanal, it is expressed by the increase of exudation and secretion,congestion and oedema of cervix mucosa, microorganism infection andetc., and it is happened in the chronic cervix inflammation. Thissymptom is not only deferred and difficult to be cured and substantiallyinfluent the normal life, but it is also possible to be developed intocancer.

[0003] At present, in China and other countries, most of the treatmentmethods of the cervical erosion use physical therapy in clinic such aselectric ironing, freezing, laser and other means to make erosive tissueto be necrotized and obscised, or use corrosive medicament such aspotassium dichromate, silver nitrate etc. to destroy the erosive tissue;for serious patients, even use surgical operation to cut out the cervixor uterus. The therapies mentioned above would bring great pains to thepatients; when removing erosive tissue, a lot of normal tissues will beinvolved. Other medicaments purchased from market generally use ethanolas solvent, so it has a strong stimulation, while the solute itself alsohas stimulation to cause pain and injure normal skin. Furthermore, mostall of these products are functionally limited to sterilization, andhave an effect to control infection, but destroy the equilibrium ofmicroorganism state, and cannot cure the infection radically. Therefore,it is necessary all along to develop a medicament for treating cervixerosion in the field.

[0004] In the research of “bio-waves” theory, the present inventor hasset up a bacterial wave growth model. Through researching, it is knownthat this wave is of its intrinsic regulation mechanism: some chemicalsubstances are able to participate the regulation in the bio-waveprocess, so as to transform an abnormal periodic slow wave into a normalphysiological chaotic quick wave, and this kind of substances are knownas promoting wave factors. Through separating, purifying andidentifying, it is determined that one of the factors isN-acetyl-D-glucosamine, the promoting wave function of which is shown inlubricating and protecting the cell. Many biochemical and physiologicalprocess of human body need the participation of the promoting wavefactors, and it would lead to an abnormal state, if this kind ofpromoting wave factors is lacked in the living body.

[0005] N-acetyl-D-glucosamine is a chemical reagent. From the 1990's, itis continually used to treat pericementitis (WO9102530A1),microbiological infection (WO9718790A3), intestinal inflammation(WO9953929A1), cornea disease (JP10287570A2), hypertrophy of theprostate (U.S. Pat. No. 5,116,615) and so on. It is also applied incosmetology (JP59013708A2), shampoo preparation (JP2011505A2), tissuegrowth regulation agent (WO/A 8 702244), and etc., but it has not beenused in the manufacture of a medicament for treating cervix erosiondisease up to now.

[0006] The applicant of the present invention finds thatN-acetyl-D-glucosamine is able to effectively treat cervix erosion, thisdiscovery is extremely beyond people's expectation, because curingcervix erosion needs many assistant cooperation of controllingmicroorganism infection, resist local exudation, removing tissueinflammation oedema and pain, and improving tissue reparation and etc.,so that doctors have to prescribe multiple medicaments combined witheach other to cure the patients, but now it is found that a medicamentwith N-actyl-D-glucosamine as active component by itself is able to curecervix erosion. Although the current techniques have disclosed theresistance for microorganism infection and tissue growth regulatingfunctions of N-actyl-D-glucosamine, the discovery of the presentapplication is quite a surprise to the people.

CONTENTS OF THE INVENTION

[0007] Therefore, the present invention is related to the use ofN-acetyl-D-glucosamine and pharmaceutical acceptable salt thereof in themanufacture of a medicament for treating cervical erosion.

[0008] In addition, the present invention is related to a method fortreating cervical erosion, including administrating to a patient who isin need thereof an effective amount of N-acetyl-D-glucosamine orpharmaceutical acceptable salts thereof.

[0009] The molecular formula of N-acetyl-D-glucosamine is C₈H₁₅NO₆, itsstructure is as follows:

[0010] N-acetyl-D-glucosamine can be purchased in the market or preparedaccording to known methods. For instance, patent application WO97/31121has disclosed a method for preparing N-acetyl-D-glucosamine from chitinby enzyme method, Japanese patent application JP63273493 has disclosed amethod in which chitin is partially hydrolyzed into N-acetyl-chitose,and then it is treated with enzyme to obtain N-acetyl-D-glucosamine.

[0011] The pharmaceutical acceptable salts of N-acetyl-D-glucosaminethat can be mentioned are the salts formed with pharmaceuticalacceptable acids, for instance, the salts formed with inorganic acids,such as hydrochloride, hydrobromide, borate, phosphate, sulfate sulfiteand hydrophosphate, and the salts formed with organic acids, such ascitrate, benzoate, ascorbate, methyl sulfate, naphthalene-2-sulfonate,picrate, fumarate, maleate, malonate, oxalate, succinate, acetate,tartrate, mesylate, tosylate, isethionate, α-ketoglutarate, α-glycerylphosphate and glucose-1-phosphate.

[0012] Generally, the compound of formula (I) of the present inventionis formulated with one or more pharmaceutically acceptable excipientsand/or carriers to prepare external-using preparations in the form ofaqueous solution (lotion), emulsion, cream, ointment, suppository, andetc, for treating cervix erosion. The amount of the active substance is0.26% of that of the whole preparation. The daily dosage ofN-acetyl-D-glucosamine is 100˜2000 mg for each person.

[0013] Though having no intention to be limited by any theories, thepresent inventor thinks that the effect of the compound of formula (I)for treating cervix erosion is affected by regulating the cellularredistribution of organism. The cellular redistribution refers to thecontinually replacement of the position of the organism cell or theposition of the microorganism cell, and rhythmic replacement of gel-solstates of biological macromolecule in vivo. Through cellularredistribution in different level, N-acetyl-D-glucosamine is able todevelop its special efficacy. The macroscopic replacements of cellularposition have the wave growth characteristic. Through regulating thewave growth of the organism cell and microorganism cell to be normal,N-acetyl-D-glucosamine makes microorganism cannot be planted locally. Inmicroorganism ecological efficacy, a method which mainly support normalbacterial colony to grow but not supplement ecological bacterial colonyis able to avoid the problem of adaptability to field planting conditionexisted in the supplementing bacterial colony. In the aspect ofrepairing skin mucosa tissue, N-acetyl-D-glucosamine has a controllingeffect for inflammation, injure, infection, exudation, and this is justthe characteristic of the product of the invention which can be widelyapplied to control the symptom and carry out the treatment to solve theradical problem.

OPTIMAL MODE FOR CARRYING OUT THE INVENTION

[0014] The following experimental examples are used to illustrate thepromoting wave function, low toxicity, effectiveness for resisting thefield planting activity of microorganism, and clinic observation fortreating cervix erosion of the compound of the present invention (thecompound of formula (I)).

[0015] I. Promoting Wave Test of the Compound of Formula (I)

[0016] 1. Experimental Materials and Method:

[0017] 1.1 Samples: Pure Compound of Formula (I)

[0018] 1.2 Experimental Materials:

[0019] Strain: Proteus Mirabilis, Lactobacilli.

[0020] Culture medium: modified LB culture medium (the components of thecomposition are: trytones of 1%, yeast extract of 0.5%, sodium chloride1%, glucose of 0.1%, TTC of 0.002% and pH=7.2˜7.4).

[0021] 1.3 Experimental Method:

[0022] The Proteus Mirabilis were inoculated at the center of LB plate,incubating at 37° C. for 9 hours, then there were concentric ringsemerged, which were extended outward continually with an interval of 3hours, and this was taken as a control; adding the compound of formula(I) with final concentration of 0.5% onto the LB plate, The ProteusMirabilis were innoculated by the same method, cultured at 37° C., andthe result showed that not only the concentric rings formed with aninterval of 3 hours were emerged, comparing with the control, it can beseen that there were also many fine waves on each ring emerged.

[0023] Meanwhile, the result of liquid cultivation also shows that, thecompound of formula (I) can improve the growth of Lactobacilli.

[0024] 2. Experimental Results and Evaluation:

[0025] The experiment adopts a bio-wave model which is used to researchthe promoting wave function of the compound of formula (I). It can beseen from the result that the compound of formula (I) was not only ableto cause bacterial cell to reveal a normal bio-wave characteristic, butalso cause the wave reveal finer wave mode, and these indicated that thecompound of formula (I) have promoting function to bio-waves, and thepromoting wave function is able to participate the reparation andre-distribution function of the cells of skin.

[0026] II. Toxicological Test of the Compound of Formula (I), including:

[0027] 1. acute toxicity test: including tests of administratingmedicine by oral, Intravenous injection and maximum limit amount foradministration;

[0028] 2. Ames test;

[0029] 3. micronucleus test of bone marrow cell of mouse;

[0030] 4. abnormal sexual test for the sperm of mouse;

[0031] 5. abnormal aberrance test for the chromosin of mouse's testis;

[0032] 6. chronic lethal test;

[0033] 7. subchronic toxicity ( feed for 90 days) test;

[0034] 8. traditional aberrance-inducing test;

[0035] The results from these tests show that in the acute toxicity testof the compound of formula (I), the dosage more than 2 g/kg is taken,which is 300 times than the injection dosage for human being, but theacute toxicosis reaction had not appeared yet; in the long-periodtoxicity test, the maximum dosage has reached up to 1 g/kg, and afterthe treatment and observation for four weeks, there is no toxicosisreaction yet; and in the reproduction test, the mouse was fed withroutine dosages from 7 mg/kg for 3 generations, it has been proved thatthe compound of formula (I) has no influence on the pregnancy, birth,nurse and the growth of baby mice, so it is proved that the compound offormula (I) is a substance without toxicity.

[0036] III. Test of Removing Pathogenic Microorganism by Resistance toField Planting Action

[0037] The results from in situ bacterial field planting, qualitativeand quantitative determination tests show that the compound of formula(I) substantially has no sterilizing or bacteria-inhibiting effect, itis by the way of disabling pathogenic microorganism such as Candidaalbicans, vagina Heamophilus and etc. from field planting growth, so asto reach the aim of resisting infection.

[0038] IV. Clinic Test

[0039] A spray formulation of 2% N-acetyl-D-glucosamine was applied tohuman beings, up to the end of August 1999, there were 123 cases ofpatients to be observed in total. It can be seen from the result,compared with blank control group (without administrating medicament),and compared with positive medicament (AgNO₃) control group, thestatistic analysis show that, the treatment group of the invention haveobvious effect in view of positive medicament control group and blankcontrol group in respect of the cure rate and effective rate, the sprayformulation of N-acetyl-D-glucosamine has a remarkable curative effectto cervix erosion, the results of which is as follows: Number of casescured effective Treating group 83 48 15 Positive medicament 60 5 8control group Blank control group 40 2 6

1. A use of N-acetyl-D-glucosamine and pharmaceutical acceptable saltsthereof in the manufacture of a medicament for treating cervicalerosion.
 2. The use according to claim 1, wherein said medicament is inthe form of aqueous solution, emulsion, cream, ointment and suppository,that for external use.
 3. The use according to claim 1 or 2, wherein thedaily dosage of said medicament is in the range of 100˜2000 mg ofN-acetyl-D-glucosamine.